Acylamino-cephem-carboxylic acids and process for preparing them

ABSTRACT

IN WHICH R1, R2 and R3 represent hydrogen or lower alkyl groups and R1 and R2 may form together an alkylene group which may be substituted, R4 represents a linear or branched alkyl radical of 1 to 5 carbon atoms, a cyclo-alkyl radical of 3 to 7 carbon atoms which may be interrupted by heteroatoms, X represents a single bond or NH, A represents a phenylene or thienylene group which may be substituted and Y represents a single bond or oxygen, and their physiologically tolerated salts; the novel compounds have very good anti-bacterial properties.   Acylamino-cephem-carboxylic acids of the general formula

United States Patent [191 Schorr et al.

[ NOV. 18, 1975 ACYLAMINO-CEPHEM-CARBOXYLIC ACIDS AND PROCESS FORPREPARING THEM Inventors: Manfred Schorr, Frankfurt am Main; ManfredWorm, Mainz; Elmar Schrinner, Wiesbaden, all of Germany Assignee:Hoechst Aktiengesellschaft,

' Frankfurt am Main, Germany Filed: May 2, 1973 Appl. No.: 356,466

Foreign Application Priority Data May 5, 1972 Germany 2222140 Apr. 3,1973 Germany 2316541 US. Cl. 260/243 C; 424/246 Int. Cl. C07D 501/24Field of Search 260/243 C References Cited UNITED STATES PATENTS 7/1973Schorr et al. 260/239.l

Primary Examiner-Nicholas S. Rizzo Attorney, Agent, or Firm-Curtis,Morris & Safford, PC.

[5 7] ABSTRACT Acylamino-cephem-carboxylic acids of the general formulaN R o COOH 19 Claims, No Drawings The present invention relates toacylamino-cephemcarboxylic acids of the general formula I C-X-A in whichR, R and R represent hydrogen or lower alkyl groups and R and R may formtogether an alkylene group which may be substituted, R represents alinear or branched alkyl radical of l to 5 carbon atoms, a cyclo-alkylradical of 3 to 7 carbon atoms which may be interrupted by hetero-atoms,Xrepresents a single bond or NH, A represents a phenylene or thienylenegroup which may be substituted and Y represents a single bond or oxygen,and to their physiologically tolerated salts.

The invention furthermore relates to a process for the preparation ofthe acylamino-cephem-carboxylic acids of the general formula I and oftheir physiologically tolerated salts, wherein a 7-amino-A -cephem-4-carboxylic acid of the general formula II S H 11 E COOH in which R hasthe meaning given above, preferably in the form of a salt or ester, isreacted with a carboxylic acid of the general formula III CX'A-YCH,COOH(Ill) in which R, R R, X, A and Y have the meanings given above, inparticular in the form of a derivative which is reactive towards anamino group, or of a salt of such a compound. The carboxyl group whichmay be protected is then freed and, if desired, the compound obtained isconverted into a physiologically tolerated salt.

If R, R and R stand for an alkyl group, there may be used in thisrespect straight chain or branched alkyl groups of l to 5 carbom atoms,the sum of the carbon atoms in the groups R, R and R being not greaterthan 6. y

R may be, for example methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, 'pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, tetrahydrofuranyl or tetrahydrothienyl.

The alkylene groups which may be formed from the groups R and R may bethose which contain 2 to 4 carbon atoms. As substituents of the alkylenegroup, there may be mentioned, for example low'molecular weight alkylgroups of l to 4 carbonatoms which them- COOH selves may also form aring that may be interrupted by a hetero-atom, preferably an oxygenatom. The alkyl substituents of the alkylene group which are not closedto a ring may also contain a hetero-atom, preferably an oxygen atom.

A stands in particular for the 1,4-phenylene or 2,5- thienylene groupand these groups may also be substituted, for example by low molecularweight l to 4 carbon atoms) alkyl, alkoxy or halogen, preferably chlo-The following compounds of the formula III may be used, for example, asstarting compounds in the process of the invention:

4-amidino-phenylacetic acid, 7

4-N-methyl-amidino-phenylacetic acid,

4-N,N-dimethyl-amidino-phenylacetic acid,4-N,N-dimethyl-amidino-phenylacetic acid,4-N,N,N-trimethyl-amidino-phenylacetic acid,4-N-ethyl-amidino-phenylacetic acid,4-N,N'-dipropyl-amidino-phenylacetic acid,4-N-i-pentyl-amidino-phenylacetic acid,4-N,N-dimethyl-N'-ethylamidino-phenylacetic acid,4-(2-imidazolinyl)-phenylacetic acid,

4-( 1,4,5,6-tetrahydro-2-pyrimidyl)-phenylacetic acid,

4-( l-methyll ,4,5 ,6-tetrahydro-2-pyrimidyl phenylacetic acid,

4-[5 ,5-bis(2-methoxy-ethyl)-1,4,5 ,6-tetrahydro-2-pyrimidyl1-phenylacetic acid,

4-( l ,5 -dimethyl-2-imidazolinyl)-phenylacetic acid,

4-( l-methyl-5-butyl-2-imidazolinyl )-phenylacetic acid,

4-( 5 ,S-dimethyl-l ,4,5 ,6-tetrahydro-2-pyrimidyl)- phenylacetic acid,

4-( l ,4,6,7,8,9-hexahydroxy-5H-cyclopentenyl[d-]pyrimidyl-2)-phenylacetic acid, 4-( 9-oxa-2,4-diazospiro[5 ,5]-undec-2-ene-3-yl phenylacetic acid,4-(2,4-diaza-spiro[5,5]undec-Z-ene-yl )-phenylacetic acid,4-(2,4-diaza-spiro[5,4]-dec-2-ene-3-yl)-phenylacetic acid,4-N,N-tetramethylene-amidino-phenylacetic acid, 3-amidino-phenylaceticacid, 4-amidino-2-methyl-phenylacetic acid,4-amidino-2-methoxy-phenylacetic acid, 4-amidino-2-butoxy-phenylacticacid, 4-amidino-2-chloro-phenylacetic acid,

'3 and the corresponding phenoxy-, thienylor thienyloxy-acetic acids orthe corresponding acids of the formula III in which X stands for NH.

As compounds of the formula, there may be for example the followingcompounds:

3-methyl-7-amino-A-ccphem-4-carboxylic acid,

3-cthyl-7-amino-A-cephem-4-carboxylic acid,3-isopropyl-7-amino-M-cephem-4-carboxylic acid.3-n-propyl-7-amino-M-cephem-4-carboxylic acid, 3-cyclopentyl-7-amino-A-cephem-4-carboxylic acid.

The carboxylic acids of the general formula III, in which X represents asingle bond, are prepared in known manner, for example from4-cyano-phenyl-, 4- cyano-phenoxyor S-cyanothienylorS-cyanothienyloxy-acetic acid esters. After conversion of the nitrilogroup into an ir'nino ether, the resulting compound is reacted withammonia or an amine or diamine to give the amidine and, finally, thecarboxylic acid ester group or an acid amide group resulting from it ishydrolyzed.

The carboxylic acid of the general formula Ill in which X stands for NHis prepared in known manner from the corresponding aminocarboxylic acidor aminocarboxylic acid derivatives by reacting the amino group of thesecompounds with cyanamides, carbodiimides, thioureas,S-alkyl-isothioureas, O-alkyl-isoureas or similar substances which maybe substituted (Houben-Weyl 8, I80 195). On the other hand, it is alsopossible to react the amino group of the abovespecified amino-carboxylicacid derivatives in known manner (Houben-Weyl 9, 887) to yieldthio-ureas or S-alkyl-isothioureas and then to react the latter withamines to obtain the carboxylic acids of the general formula ll in whichX stands for NH.

The 7-amino-A -cephem-4-carboxylic acids of the formula II usedaccording to the invention as starting materials can be prepared in amanner known from literature or according to French Patent applicationNos. 71/47761 or 72/31699.

The novel acyl derivatives of the 7-amino-A -cephem-4-carboxylic acid ofthe formula II are obtained in a particularly advantageous manner byreacting the 7- amino-A -cephem-4-carboxylic acid of the formula II inthe form of its alkali metal salt or tertiary amine salt, for examplethe sodium, potassium or triethylamine salt, with a reactive derivativeof a carboxylic acid of the general formula III in an as far as possibleneutral pH-range, preferably in the pH-range of from 6 to 9.

The salts of the 7-amino-A -cephem-4-carboxylic acid of the formula [Imay be used directly or they may prepared in the solution used for thereaction from 7- amino-A -cephem-4-carboxylic acid of the formula [Iand, for example, sodium bicarbonate, di-sodiu'mhydrogen-phosphate ortriethylamine. I

The 7-amino-A -cephem-4-carboxylic acid of the formula ll may also beused in the form of its derivatives such as, in particular, its esters;in this case those esters are preferred which can be cleaned underrelatively mild conditions, for example by hydrogenolysis or byhydrolysis, for example with trifluoroacetic acid, for example thebenzhydryl ester, benzyl ester, 4 methoxybenzyl ester, 4-nitro-phenylester, tert. butyl ester or trichloroethyl ester. 7

in some cases, the reactive derivatives may not only be used in the formof a hydrochloride, but also in the form of other salts obtained in theprocess of preparation.

used,

In general, the reaction according to the invention is carried out inwater. However, it is also possible to operate in the presence ofsolvents which are miscible with water, for example acetone,dimethylformamide, dimethylacetamide, dioxane or tetrahydrofurane.

For the acylation of the 7-amino-A -cephem-4-carboxylic acid of theformula II according to the invention, in particular the acid chlorideshave proved to be good as reactive derivatives of the carboxylic acidsof the general formula III. They can be obtained in known manner fromthe carboxylic acids by the action of thionyl chloride in an inertsolvent, for example an aromatic hydro-carbon. In such a case they areobtained in the form of hydrochlorides which can directly be used forfurther reaction. Besides the acid chlorides, also other reactivederivatives of the carboxylic acids of the formula III may be used, forexample acid bromides, activated esters such as the p-nitrophenyl-ester,pnitrophenyl-thio-ester or cyanomethyl ester, acid azides'or symmetricor mixed anhydrides. Reactive addition compounds of the carboxylic acidsof the general formula [II and, for example, carbodiimides that act ascondensation agents may likewise be used, the 7- amino-A-cephem-4-carboxylic acid of the formula II being advantageously usedwith a protected carboxyl group, for example in the form ofa salt or ofan ester. The reactive derivatives may be used not only in the form ofthe hydrochloride, but also in the form of other salts obtained duringthe preparation.

The acylation of the 7-amino-A -cephem-4-carboxylic acid of the formulaII is advantageously carried out by adding equi-molar amounts or aslight excess amount of a compound of the formula Ill in a form of areactive derivative to the solution of a salt of the 7- amino-A-cephem-4-carboxylic acid of the formula II in water or in a mixture ofwater and a solvent which is miscible with water. 7

It has been found to be particularly advantageous to introduce thehydrochloride of the acid chloride in solidform. in order to bind thehydrogen chloride, the solution is previously combined with twoequivalents of a base, for example sodium bicarbonate or triethylamine.The reaction is carried out at room temperature or slightly reducedtemperatures, preferably between 5 and +5 C. the pH-value which is atfirst weakly alkaline falls during the reaction to about 7.

The basic cephem-carboxylic acids of the general formula I are obtainedin the form of inner salts and crystallize in most cases and can thenbe=,isolated by filtration with suction. In some other cases, it isnecessary to precipitate themwith the aid of non-solvents, for exampleacetone, and for purification they must be recrystallized from suitablesolvent mixtures or again dissolved and recrystallized.

The novel acylamino-cephem-carboxylic acids of the general formula Icontain in their molecule an amidino or guanidino group which may besubstituted and have, therefore, an amphoteric character. They forminner salts and are generally well soluble in water with a pH- value ofabout 5.

The products of the invention may be used as such, in the form of theirphysiologically tolerated salts or also in the form of theabove-described crude products. As substances which may be used for theformation of physiologically tolerated salts, there may be mentioned,for example hydrochloric acid, hydrobromic acid, amidosulfonic acid,citric acid, acetic acid, and suitable bases such as sodium hydroxide,ammonia or ethanolamine. The salt formation is effected, for example, bymixing equivalent amounts in solution and concentrating by evaporation.

The novel acylamino-cephem-carboxylic acids have outstandinganti-bacterial properties. Their action 5 the treatment of bacterialinfections. They can be used against gram-positive germs can be comparedwith that as such or together with the usual pharmaceutical exofpenicillins. In addition, they have the advantage of cipients andadjuvants, for example tragacanth. lactose, having a good activityagainst penicillinase-forming talc, agar-agar, etc. in the form ofgalenic preparations, staphylococci. for example tablets, dragees,capsules. or in the form of Minimum inhibition concentration in -y/ml.solutions or suspensions, which contain the active sub- Germ: Staph.aur. 285 stance in a quantity of from about 50 to 1000 mg. preferably100 500 mg. For parenteral administration, substance MIC preferablysuspensions or solutions in water are used. PenicillimGNA I25 Otherphysiologically tolerated organic solvents. for exp i im .v.l( 135 ampleethanol, polyglycols, or solubillzers may also be Ampicillin added. Itis also possible to combine them with other ff 2 active substances.Thus, the compounds of the inven- 3 12.5 tion may be appliedsimultaneously, for example, with g s? other antibiotic agents, forexample those of the series 6 I1 of the penicillins, cephalosporins, orwith compounds g which have a favorable influence on the symptomatol- Hogy of bacterial infections, for example antipyretic, an-

12 algesic or antiphlogistic agents. :3 3:; The following Examplesillustrate the invention. The 15 3.1 25 novelacylamino-cephem-carboxylic acids described in :3 these Examples werecharacterized by their thin layer |g chromatogram. As the layer, silicagel (Merck) was used and, a 0.l-N aqueous ammonium acetate solution wasused as the eluant. The plates were developed by The novelacylamino-cephem-carboxylic id h the action of iodine vapor. Theproducts were free from a considerably higher stability, in particulartowards unfeactfid in0-A-cephem-4-carboxylic acid used acids, thancommercially available eni i]lin in each respective case. In theinfrared spectrum, the compounds showed the absorption band at 1755 cm TpH L7 Time 24 hours which is characteristic for the B-lactam ring.

Concentration 0.05% in 0.02-N aqueous hydrochloric acid SubstanceDestroyed proportion in "/1 EXAMPLE 1 Penicillin-G-Na 100 :73-Methyl-7-(4-amidino-phenylacetamino)-A -cephemgfigg g 4-carboxylicacid ff l 40 a. 5.35 g of 4-amidino-phenylacetic acid (melting 5 6 "Apoint 295 297 C, decomposition) were suspended in 8:; ml of anhydrousbenzene. Two drops of dimethyl- 6 5 ("A formamide and 17.9 g ofthionylchloride were added 7 l0 and the whole was heated for 1 /2 hoursunder reflux. 8 3 45 After having cooled, the solid product was filteredoff with suction, washed with anhydrous benzene and dried under reducedressure. 6.9 of 4-amidinowhen Compared wlth the cephalosporms avallablephenylacetic acid ch ioride-hydroch loride, melting 9 point 174 177 C(decomposition). were obtained. aclds Sh0w.wlth slmllar acne? againstgram-posmve b. 3.64 g of 4-amidino-phenylacetic acid chloridef si,pharmacokmeuc properties: "9" hydrochloride were added within about 10minutes to a ll'lIrZlVfiflOUSTnjECtlOll of the substances of theinvention Solution which had been cooled to about C of 278 g 1nto dogs,higher and longer lasting substance levels in of 3 methyl 7 amino A3cephem 4 carboxylic acid and glzffi z g f 3.36 g Of sodium bicarbonatein 50 ml of water and 6 ml of acetone. The reaction mixture foamed u anda Test Subtlhs ATCC 6633 colorless crystalline precipitate was formedwhich was Serum, filtered off with suction at 0 C after 1 hour, washed$3 ?A f :22: ii f gyi f g s iigggggg with cold water, acetone and etherand dried in air.

' 5.5 g of 3-methyl-7-(4-amidino-phenylacetamino)- Preparation FSC' Ah 1h 2h 3h 4h 5h 6h T5 Cephalotin 14.5 10.4 7.4 3.7 1.9 0 0 0 1.0 Example 435.2 30.2 23.3 20.5 7.3 6.5 5.7 3.4 1.8 Example 5 25.5 21.3 17.8 12.58.7 6.1 4.3 3.0 1.9

The values indicated in the above Tables show that the novelacylamino-cephem-carboxylic acids are 6 equivalent to the antibioticsavailable in commerce and that in special cases they are even superiorto them.

Thus, the products of the invention are valuable therapeutic agentswhich can be used very successfully for A -cephem-4-carboxylic acid,melting point 230 233 C (decomposition), were obtained.

EXAMPLE 2 3-Methyl-7-( 4-amidino-phenoxyacetamino)-A"-cephem-4-carboxylic acid a. 9.7 g of 4-amidino-phenoxyacetic acid(melting point 324 326 C) were suspended in 50 ml of anhydrous benzene,29.7 g of thionyl chloride were added and the whole was heated for 1hour on the steam bath under reflux. After having allowed the reactionmixture to cool, the solid product was filtered off with suction. l 1.4g of 4-amidino-phenoxy-acetyl chloride-hydrochloride were obtained whichhad a decomposition point of 142 143 C.

b. From this compound, there was obtained in a manner analogous to thatdescribed in Example 1, the 3-methyl-7-(4-amidino-phenoxyacetamino)-A-cephem- 4-carboxylic acid in theform of colorless crystals which decompose at'temperatures starting from240 C nwards.

EXAMPLE 3 3-Methyl-7-( 5-amidino-2-thienylacetamino)-A-cephem-4-carboxylic acid In a manner analogous to that described inExample 1, there was obtained from 5-amidino-thienyl-Z-acetic acid and3-methyl-7-amino-A-cephem-4-carboxylic acid, the3-methyl-7-(5-amidino-2-thienylacetamino)- A -cephem-4-carboxylic acidin theform of weakly yellowish crystals which were found to melt atabout 225 C with decomposition.

EXAMPLE 4 3-Methyl-7-[4-( 2-imidazolinyl )-phenylacetamino]-A-cephem-4-carboxylic acid la. g of 4-(2-imidazolinyl)-pheny-lacetic acid(melting point 196 197 C) were stirred with 150 ml of anhydrous benzeneand 90 ml of thionyl chloride for 5 hours at 50 60 C. After cooling, thecrystalline product was filtered off with suction, washed with anhydrousbenzene and dried under reduced pressure. 1 1 g of4-(2-imidazolinyl)-phenylacetic acid chloridehydrochloride wereobtained.

b. 1.81 g of 4-(2-imidazolinyl)-phenylacetic acid chloride-hydrochloridewere added within about 10 minutes to a solution which had been cooledto about 0 C of 1.29 g of 3-methyl-7-aminoA -cephem-4-carboxylic acidand 1.68 g of sodium bicarbonate in 27 ml of water and 3 m] of acetone.After 1 hour, the mixture was filtered and the pH-value thereof wasadjusted to 1.7 by means of 6N-hydrochloric acid, whereupon a jelly-likeproduct formed. The latter was filtered off with suction, washed withacetone, dissolved in 3 ml of water and neutralized with sodium acetate.The product that had then crystallized was filtered off with suction andwashed with ice-water, acetone and ether. 1.6 g of3-methyl-7-[4-(2-imidazo1inyl)-phenylacetamino]- A -cephem-4-carboxy1icacid melting at 235 237 C were obtained. The product contained one halfmole of acetone. The infrared and NMR spectra correspond to the assumedstructure.

lla. 1.24 g of N,N-dicyclohexyl-carbodiimide in 10 ml of DMF were addeddropwise to a solution that had been cooled to 0 C of 1.9 g of3-methyl-7-amino-A cephem-4-carboxylic acid-benzhydryl ester (meltingpoint 153 154 C) and 1.2 g of 4-(2-imidazolinyl)- phenylacetic acidhydrochloride (melting point 223 8 224 C) in 50 ml of DMF. After havingallowed the whole to stand for 1 hour at 0 C and overnight at 20 C. themixture was filtered and the product was precipitated from the filtratewith the aid of ether; it was obtained in the form of an oil. This oilwas crystallized with acetone and recrystallized from ethanol.

In this manner, 3-methyl-7-[4-imidazolinyl)- phenylacetaminol-A-4-carboxylic acid benzhydryl ester-hydrochloride in the form ofcolorless crystals which were found to melt at about 200 C withdecomposition was obtained.

The infrared spectrum showed the characteristic bands to be expected:

B-Lactam: 1770, ester: 1715, and acyl: 1660 cm".

b. The ester obtained according to Example 4 II(a) was suspended in 2.5ml of benzene and 0.2 ml of anisole and the whole was treated for 15minutes at room temperature with 0.3 ml of trifluoroacetic acid. Thevolatile components were distilled off under reduced pressure and theresidue was washed with ethyl acetate. The residue was dissolved in 10ml of water, the solution was filtered and the pH-value was adjusted to7 by means of dilute ammonia. After the addition of a small amount ofacetone and cooling, the 3-methyl-7-[4-(2-imidazolinyl)-phenylacetamino]-A -cephem-4-carboxylic acid crystallized.It was filtered off with suction and washed with a mixture of acetoneand water. The colorless crystals were found to melt at 235 C.

EXAMPLE 5 3-Methyl-7-[4-( l ,4,5,6-tetrahydro-2-pyrimidyl)-phenylacetamino]-A -cephem-4-carboxylic acid In a manner analogous tothat described in Example 4, there was obtained from4-(1.4,5,6)-tetrahydro-2- pyrimidyl-phenylacetic acid hydrochloride(melting point 256 257 C with decomposition) and 3-methyl- 7-amino-A-cephem-4-carboxylic acid, the 3-methyl-7-[4-(1,4,5,6-tetrahydro-2-pyrimidyl)-phenylacetamino]-A-cephem-4-carboxylic acid melting at 243 244 C (decomposition).

The product was recrystallized from a mixture of water and acetone andcontained then one half mole of acetone and one half of crystal water.The infrared and NMR spectrums corresponded with the expected structure.

EXAMPLE 6 3-Methyl-7-[4-( Z-imidazolinyl)-phenoxyacetamino]- A-cephem-4-carboxylic acid a. 25 ml of thionyl choride were added to asuspension of 4.4 g of 4(-2-imidazolinyl)-phenoxyacetic acid (meltingpoint 278 280 C, decomposition) in 25 ml of anhydrous benzene and thewhole was heated for 3 hours under reflux. After having allowed thereaction mixture to cool, the solid product was filtered off withsuction and dried under reduced pressure. 4.9 g of 4- (Z-imidazolinyl)-phenoxyacetyl chloride-hydrochloride were obtained; the compound wasfound to decompose at a temperatures from 195 C onwards.

b. 4.35 g of 4-(2-imidazolinyl)-phenoxyacetyl chloride-hydrochloridewere added within about 10 minutes to a solution that had been cooled toabout 0 C of 3.05 g of 3-methyl-7-amino-A -cephem-4-carboxylic acid and3.98 g of sodium bicarbonate in 65 ml of water and 7.5 ml of acetone.After one hour, the pH-value was adjusted to 4 by means ofZN-hydrochloric acid, the mixture was filtered, the solvent wasdistilled off under reduced pressure. the residue was dissolved in hotmethanol. filtered and the product was precipitated with the aid ofacetone. 2.9 g of 3-methyl-7-[4-(2-imidazolinyl)-phenoxyacetamino]-A"-cephem-4-carboxylic acid wereobtained which were found to decompose at 202205 C (Rf 0.23).

EXAMPLE 7 3-Methyl-7-[4-( 1,4,5 .6-tctrahydro-2-pyrimidylphenoxyacetamino1-A*-cephem-4-carboxylic acid In a manner analogous tothat described in Example 6, there was obtained from4-(1,4,5,6-tetrahydro-2- pyrimidyl)-phenoxyacetic acid-hydrochloride(melting point 245 246 C) and 3-methyl-7-amino-A -cephem-4-carboxylicacid. the3-methy1-7-[4-(1.4.5.6-tetrahydro-2-pyrimidyl)-phenoxyacetamino]-A-cephem-4-carboxylic acid in the form of an amorphous powder, which was uniformin thin layer chromatography (R, 0.22).

EXAMPLE 8 3-Methyl-7-(4-guanidino-phenylacetamino )-A-cephem-4-carboxylic acid 4.7 g of 4-guanidino-phenylacetylchloride-hydro chloride (melting point 126 128 C) were added withinabout 10 minutes to a solution that had been cooled to about C. of 3.1 gof 3-methyl-7-amino-A cephem-4-carboxylic acid and 4.0 g of sodiumbicarbonate in 52 ml of water and 19 ml of tetrahydrofurane. Afterminutes at 0 C and 10 minutes at room temperature, the whole wasfiltered, the product was precipitated from the filtrate by means ofacetone and recrystallized from a mixture of water and acetone. 3.1 g of3-methy1-7-(4-guanidino-phenylacetamino)-A cephem-4-carboxylic acidmelting at about 270 C (decomposition) were obtained.

EXAMPLE 9 3-Methyl-7-( 4-N,N -tetramethylene-amidinophenylacetamino)-A-cephem-4-carboxylic acid a. 1.24 g of N,N'-dicyclohexyl-carbodiimide in10 ml of DMF were added dropwise to a solution that had been cooled to 0C of 1.9 g of 3-methyl-7-amino-A cephem-4-carboxylic acid benzhydrylester (melting pont 153 154 C) and 1.34 g of4-N,N-tetramethylene-amidino-phenylacetic acid hydrochloride (meltingpoint 225 228 C with decomposition) in 50 ml of DMF. After havingallowed the whole to stand for 1 hour at 0 C and overnight at about 20C, it was filtered and from the filtrate the product was precipitatedwith the aid of ether, crystallized with a small amount of water andacetone and recrystallized from ethanol.

In this manner, 3-methyl-7-(4-N,N'-tetramethy1ene-'amidinophenylacetamino)-A -cephem-4-carboxylic acid ester hydrochloridewas obtained in the form of colorless crystals which were formed to meltat about 175 C with decomposition.

The infrared spectrum showed the expected characteristic bands:

B-lactam: 1770, ester: 1715, acryl: 1630 cm.

b. The ester obtained according to Example 9(a) was suspended in 5 ml ofbenzene and 0.5 ml of anisole and treated for minutes at roomtemperature with 0.75 ml of trifluoroacetic acid. The volatilecomponents were then removed by distillation under reduced pressure andthe residue was washed with ethyl acetate.

This residue was dissolved in 10 ml of water and 1 ml of acetone, thesolution was filtered and the pP-value thereof was adjusted to 7 bymeans of dilute ammonia. After having added a small amount of acetoneand having allowed the mixture to cool, the 3-methyl-7-(4-N.N-tetramethylene-amidino-phenylacetamino)-A- cephem-4-carboxy1ic acidcrystallized. It was filtered off with suction and washed with a mixtureof acetone and water. The colorless crystals were found to melt at about212 C with decomposition.

EXAMPLE l0 3-Methyl-7-[4-(N-methyl-1,4,5 ,6-tetrahydro-2- pyrimidyl)-phenoxyacetamino]-A -cephem-4-carboxylic acid. 7

In a manner analogous to that described in Example 6, there was obtainedfrom 4-(N-methyl-l ,4,5,6-tetrahydro-2-pyrimidyl)-phenoxyacetic acid and3-methyl-7-amino-A -cephem-4-carboxylic acid, the 3-methyl-7-[4-N-methyl-l ,4,5 ,G-tetrahydro-2-pyrimidylphenoxyacetamino]-A-cephem-4-carboxylic acid in the form of colorlesscrystals that were found to melt at about 250 C with decomposition (R0.34).

EXAMPLE l1 3-Ethyl-7-(4-guanidino-phenylacetamido)-A -cephem-4-carboxy1ic acid 0.59 g of 4-guanidino-phenylacetylchloride-hydrochloride were added within about 10 minutes to a solutionthat had been cooled to about 0 C of 0.46 g of 3- ethyl-7-amino-A-cephem-4-carboxy1ic acid and 0.42 g of sodium bicarbonate in 8 ml ofwater and 0.8 ml of acetone. With foaming, a fine precipitate was formedwhich was allowed to stand for 1 hour at 0 C, filtered off with suction,washed with a small amount of cold water, acetone and ether and dried inair. 0.50 g of 3- ethyl-7-(4-guanidino-phenylacetamido)-A -cephem-4-carboxylic acid decomposing at temperatures from 250 C onwards wereobtained.

EXAMPLE 12 3-Ethyl-7-[ 4-(2-imidazolinyl )-phenylacetamido ]-Acephem-4-carboxylic acid In a manner analogous to that described inExample 1 1, there were obtained from 0.62 g of 4-(2-imidazolinyl)-phenyl-acetyl chloride-hydrochloride and 0.46 g of3-ethy1-7-amino-A -cephem-4-carboxylic acid, 0.54 g of3-ethyl-7-[4-(2-imidazolinyl)- phenylacetamido]-A -cephem-4-carboxylicacid, which 'was found to melt at about C with decomposition.

EXAMPLE l3 3-Ethy1-7-[4-( N-methyl-l ,4,5 ,6-tetrahydro-2- pyrimidyl)-phenoxy-acetamido -A -cephem-4-carboxylic acid In a manner analogousto that described in- Example 11, there were obtained from 0.73 g of4-(N-methyll,4,5,6-tetrahydro-2-pyrimidyl)-phenoxyacetylchloride-hydrochloride and 0.46 g of 3-ethyl-7-amino-Acephem-4-carboxylic acid, 0.87 g of 3-ethyl-7-[4-(N-methyl-1,4,5,6-tetra-hydro-2-pyrimidy1)-phenoxyacetamido[-A-cephem-4-carboxylic acid was obtained which was found to melt at 235237 C with decomposition.

EXAMPLE l4 3-Isopropyl-7-[4-(Z-imidazolinyl)-phenylacetamido]- A-cephem-4-carboxylic acid In a manner analogous to that described inExample 11, there were obtained from 0.13 g of 4-(2-imidazolinyl)-phenylacetylchloride, hydrochloride and 0.09 g of3-isopropyl 7-amino-A -cephem-4-carboxylic acid, 0.15 g of3-isopropyl-7-[4-( 2-imidazolinyl)- phenylacetamidol-A-cephem-4-carboxylic acid which was found to melt at 201 203 C withdecomposition.

EXAMPLE l5 3-Ethyl-7-(4-amidino-phenylacetamido)-A -cephem 4- carboxylicacid In a manner analogous to that described in Example 1], there wereobtained from 0.395 g of 4-amidinophenylacetyl-chloride-hydrochlorideand 0.385 g of 3- ethyl-7-amino-A -cephem-4-carboxylic acid, 0.60 g of3-ethyl-7-(4-amidino-phenylacetamido)-A -cephem-4- carboxylic acid whichwas found to melt at 224 226 C with decomposition.

EXAMPLE l6 3-Ethyl-7-(4-amidino-phenoxyacetamido)-A -cephem-4-carboxylic acid In a manner analogous to that described in Example ll,there were obtained from 0.50 g of 4-amidin0-phenoxyacetyl-chloride-hydrochloride and 0.455 g of 3-ethyl-7-amino-A-cephem-4-carboxylic acid, 0.610 g of3-ethyl-7-(4-amidino-phenoxy-acetamido)-A -cephem-4-carboxylic acidwhich was found to melt at 239 240 C with decomposition.

EXAMPLE l7 3-Isopropyl-7-( 4-amidino-phenylacetamido)-A-cephem-4-carboxylic acid In a manner analogous to that described inExample 11, there were obtained from 0.40 g of4-amidinophenylacetyl-chloride-hydrochloride and 0.36 g of 3-isopropyl-7*amino-A -cephem-4-carboxylic acid, 0.57 g of3-isopropyl-7-(4-amidino-phenylacetamido)-A cephem-4-carboxylie acidwhich was found to melt at 233 235 C with decomposition.

EXAMPLE l8 We claim: 1. An acylamino-cephem-carboxylic acid of theformula COOH

and physiologically tolerated salts thereof, wherein R, R and R takenalone, are hydrogen'or lower alkyl; R and R taken together, are alkylenehaving 2 to 4 carbon atoms,alkylene having from 2 to 4 carbon atomssubstituted with lower alkyl or lower alkoxy alkyl, or alkylene havingfrom 2 to 4 carbon atoms substituted on one carbon atom thereof withlower alkylene or lower oxa-alkylene to form a Spiro-connected 5- or 6-membered carbocycle or oxygen heterocycle therewith; R is linear orbranched alkyl having I to 5 carbon atoms, cycloalkyl having 3 to 7carbon atoms, tetrahydrofuranyl, or tetrahydrothienyl; X is NH or asingle bond; A is phenylene, thienylene, or phenylene or thienylenesubstituted by lower alkyl, lower alkoxy, or chlorine; and Y is oxygenor a single bond.

2. 3-Methyl-7-(4-amidino-phenylacetamino)-A cephem-4-carboxylic acid.

3. 3-Methyl-7-(4-amidino-phenoxyacetamino)-A cephem-4-carboxylic acid.

4. 3-Methyl-7-(5-amidino-2-thienylacetamino)-A cephem-4-carboxylic acid.

5. 3-Methyl-7-[4-(2-imidazolinyl)- phenylacetamino]-A-cephem-4-carboxylic acid.

6. 3-Methy|-7-[4-( l ,4,5,6-tetrahydro-2-pyrimidyl)- phenylacetaminol-A-cephem-4-carboxylic acid.

7. 3-Methyl-7-[4-( 2-imidazolinyl)-phenoxyacet aminol-A-cephem-4-carboxylie acid.

8. 3-Methyl-7-[4-( l,4,5,6-tetrahydro-2-pyrimidyl)- phenoxyacetamino]-A-cephem-4-carboxylic acid.

9. 3-Methyl-7-(4-guanidino-phenylacetamino)-A cephem-4-carboxylic acid.

l0. 3-MethyI-7-(4-N,N'-tetramethylene-amidinophenylacetamino)-A-cephem-4-carboxylic acid.

1 l. 3-Methyl-7-[4-( N-methyll ,4,5 ,6-tetrahydro-2-pyrimidyl)-phenoxyacetaminol-A -cephem-4-carboxylic acid.

12. 3-Ethyl-7-(4-guanidino-phenylacetamido)-A cephem-4-carboxylic acid.

13. 3-Ethyl-7-[4-(2-imidazoliny])-phenylacetamido]- A-cephem-4-carboxylic acid.

14. 3-Ethyl-7-[4-( N-methyl-l ,4,5 ,6-tetrahydro-2-pyrimidyl)-phenoxyacetamido]-A -cephem-4-carboxylic acid.

15. 3-Isopropyl-7-[4'-(2-imidazolinyl)- phenylacetamido]-A-cephem-4-carboxylic acid.

16. 3-Ethyl-7-(4-amidino-phenylacetamido)-A cephem-4-carboxylic acid.

17. 3-Ethyl-7-(4-amidino-phenoxyacetamido)-A cephem-4-carboxylic acid.

' l8. 3-Isopropyl-7-(4-amidino-phenylacetamido)-A cephem-4 -carboxylicacid.

19. 3-Isopropyl-7-(4-amidino-phenoxyacetamido)- A -cephem-4-carboxy1icacid.

1. AN ACYLAMINO-CEPHEM-CARBOXYLIC ACID OF THE FORMULA 2.3-Methyl-7-(4-amidino-phenylacetamino)- Delta 3-cephem-4-carboxylicacid.
 3. 3-Methyl-7-(4-amidino-phenoxyacetamino)- Delta3-cephem-4-carboxylic acid. 4.3-Methyl-7-(5-amidino-2-thienylacetamino)- Delta 3-cephem-4-carboxylicacid.
 5. 3-Methyl-7-(4-(2-imidazolinyl)-phenylacetamino)- Delta3-cephem-4-carboxylic acid. 6.3-Methyl-7-(4-(1,4,5,6-tetrahydro-2-pyrimidyl)-phenylacetamino)- Delta3-cephem-4-carboxylic acid. 7.3-Methyl-7-(4-(2-imidazolinyl)-phenoxyacetamino)- Delta3-cephem-4-carboxylic acid. 8.3-Methyl-7-(4-(1,4,5,6-tetrahydro-2-pyrimidyl)-phenoxyacetamino)- Delta3-cephem-4-carboxylic acid.
 9. 3-Methyl-7-(4-guanidino-phenylacetamino)-Delta 3-cephem-4-carboxylic acid. 10.3-Methyl-7-(4-N,N''-tetramethylene-amidino-phenylacetamino)-Delta3-cephem-4-carboxylic acid. 11.3-Methyl-7-(4-(N-methyl-1,4,5,6-tetrahydro-2-pyrimidyl)-phenoxyacetamino)-Delta 3-cephem-4-carboxylic acid. 12.3-Ethyl-7-(4-guanidino-phenylacetamido)- Delta 3-cephem-4-carboxylicacid.
 13. 3-Ethyl-7-(4-(2-imidazolinyl)-phenylacetamido)- Delta3-cephem-4-carboxylic acid. 14.3-Ethyl-7-(4-(N-methYl-1,4,5,6-tetrahydro-2-pyrimidyl)-phenoxyacetamido)-Delta 3-cephem-4-carboxylic acid. 15.3-Isopropyl-7-(4-(2-imidazolinyl)-phenylacetamido)- Delta3-cephem-4-carboxylic acid.
 16. 3-Ethyl-7-(4-amidino-phenylacetamido)-Delta 3-cephem-4-carboxylic acid. 17.3-Ethyl-7-(4-amidino-phenoxyacetamido)- Delta 3-cephem-4-carboxylicacid.
 18. 3-Isopropyl-7-(4-amidino-phenylacetamido)- Delta3-cephem-4-carboxylic acid. 19.3-Isopropyl-7-(4-amidino-phenoxyacetamido)- Delta 3-cephem-4-carboxylicacid.